Complete restoration of normal DNA repair characteristics in group F xeroderma pigmentosum cells by over-expression of transfected XPF cDNA.
XP-F cDNA was cloned into a mammalian expression vector plasmid, and introduced into group F xeroderma pigmentosum (XP-F) cells. Several cell clones possessing transfected XPF cDNA were randomly isolated, and DNA repair characteristics of a clone, XP-FR2, were extensively analyzed. The XP-FR2 cells expressed high level of XPF protein as well as ERCC1 protein, although their parental XP-F cells expressed extremely low level of both proteins. The XP-FR2 cells showed UV resistance comparable to normal human cells, and had normal levels of UV-induced unscheduled DNA synthesis and normal capability to remove cyclobutane pyrimidine dimers and (6-4) photoproducts. Frequencies and types of UV-induced mutations examined by shuttle vector plasmids in XP-FR2 cells were similar to those in normal human cells. These results demonstrate that excision repair defect in XP-F cells is fully corrected by over-expression of XPF cDNA alone, although only partial correction of the cells by XPF cDNA has been reported before.
Yagi
T
T
Department of Radiation Genetics, Graduate School of Medicine, Kyoto University, Japan. C51845@sakura.kudpc.kyoyo-u.ac.jp
Matsumura
Y
Y
Sato
M
M
Nishigori
C
C
Mori
T
T
Sijbers
A M
AM
Takebe
H
H
D001483:Base Sequence; D002472:Cell Transformation, Viral; D002478:Cells, Cultured; D004260:DNA Repair; D004268:DNA-Binding Proteins / Q000096:biosynthesis / Q000235:genetics / Q000502:physiology*; D005260:Female; D005347:Fibroblasts; D005786:Gene Expression Regulation / Q000528:radiation effects; D016153:Genes, Suppressor; D006801:Humans; D008969:Molecular Sequence Data; D016296:Mutagenesis; D017354:Point Mutation; D012343:RNA, Transfer / Q000096:biosynthesis; D011994:Recombinant Proteins / Q000096:biosynthesis; D012867:Skin / Q000378:metabolism; D014162:Transfection; D014466:Ultraviolet Rays; D014983:Xeroderma Pigmentosum / Q000235:genetics* / Q000378:metabolism
D004268:DNA-Binding Proteins; D011994:Recombinant Proteins; C096275:supF tRNA; C095888:xeroderma pigmentosum group F protein; D012343:RNA, Transfer